Methods and assays for immune phenotyping

A method of screening a test therapeutic agent comprising:

• a. providing or having been provided an immune cell;
• b. optionally determining if the immune cell has an immunosuppressive or hyper-inflammatory endotype;
• c. contacting the immune cell with a test therapeutic agent;
• and
• d. determining if one or more cytokines associated with cellular immunity are increased, decreased, or the same compared to a control or compared to before the immune cell was contacted with the test therapeutic agent.

A method of evaluating drug efficacy by measuring immune function in a subject:

• a. providing or having been provided a biological sample comprising whole blood or diluted whole blood or isolated peripheral blood mononuclear cells (PBMCs);
• b. quantitating T cell interferon-gamma (IFN-) and monocyte TNF-α production using ELISpot in the biological sample comprising whole blood or diluted whole blood;
• c. optionally determining that a subject has an immunosuppressive endotype if T cell cytokine or monocyte cytokine production is decreased compared to a control;
• and
• d. administering a drug to the subject and determining the immune function of the subject in response to the drug.

A method of reversing lymphopenia or improving T cell function in a subject comprising:

• a. providing or having been provided a biological sample from the subject;
• b. stimulating a T cell or monocyte cell or both to secrete a cytokine associated with cellular immunity;
• c. quantitating at least one cytokine associated with cellular immunity using ELISpot assay or FluoroSpot assay in the biological sample;
• and
• d. administering an immune-stimulating agent, optionally IL-7, GM-CSF, anti-PD-1, anti-PD-L1, and OX-40 agonistic Abs.